Rp-8-AHA-cAMPS-Agarose

cyclic AMP antagonist immobilized on agarose gel

BLG-A012-06

0.6 ml

257.00 USD

BLG-A012-25

2.5 ml

801.00 USD

BLG-A012-60

6 ml

1,559.00 USD

The metabolically stable protein kinase A inhibitor Rp-cAMPS immobilized on agarose by an aminohexylamino spacer attached to position 8 of the ligand. Free Rp-cAMPS binds to the protein kinase A holoenzyme but prevents dissociation to free catalytic and regulatory subunits. In addition, it is not hydrolized by mammalian phosphodiesterases.The gel can be used for affinity chromatography of various cyclic nucleotide-responsive proteins such as protein kinase A (presumably as its holoenzyme), phosphodiesterases and others.

After equilibration with about 10 column volumes of starting buffer the affinity column is loaded with the protein solution at 50-100µl/min. In order to elute unspecifically bound nucleotide-dependent proteins the column can be washed, e.g. with 1mM 5'-AMP. Elution of the target protein is performed by a cyclic nucleotide gradient up to 1mM. For elution of phosphodiesterases it could be advisable to use the highly resistant Sp-cAMPS or the completely stable Rp-cAMPS. For protein kinase holoenzyme elution the gradient should be made with Rp-cAMPS.Suitable buffer systems for your special application have to be tested but phosphate should be not optimal since one essential affinity interaction of cyclic nucleotides towards their target receptors is the cyclic phosphate.Regeneration can be achieved by a combination of up to 100 mM cAMP and buffer salts or 8 M urea.